Abstract
Cartilage erosion in degenerative joint diseases leads to lameness in affected horses. It has been reported that andrographolide from Andrographis paniculata inhibited cartilage matrix-degrading enzymes. This study aimed to explore whether this compound protects equine cartilage degradation in the explant culture model and to determine its effect on matrix metalloproteinase-2 (MMP-2) expression, a matrix-degrading enzyme, in equine chondrocyte culture. Equine articular cartilage explant culture was induced by 25 ng/mL interleukin-1β, a key inducer of cartilage degeneration, in cultures with or without andrographolide ranging from 10 to 50 μM. After 3–21 days, they were analyzed for the markers of cartilage degradation. It was found that interleukin-1β increased the release of sulfated glycosaminoglycans and hyaluronan from the explants into the culture media consistently with the decrease in uronic acid and collagen content in the cartilage explants. These catabolic effects were inhibited when cotreated with interleukin-1β and andrographolide. In primary equine chondrocytes, andrographolide suppressed interleukin-1β-induced MMP-2 mRNA expression and MMP-2 activity in the culture medium. These results confirmed the in vitro potent chondroprotective activities of this compound which were performed in cartilage explants and on a cellular level. These may indicate the application of andrographolide for therapeutic use in equine degenerative joint diseases.
Highlights
Degenerative joint disease (DJD), often called osteoarthritis, is characterized by the progressive and permanent degradation of articular cartilage [1]
Cytotoxicity assays were conducted in order to evaluate whether andrographolide exerts cytotoxicity effect on equine chondrocytes which performed in the conditions of primary cell culture and cartilage explant culture systems
On the cellular level of equine chondrocyte, we demonstrated the suppressive effect of andrographolide on expression of matrix metalloproteinase-2 (MMP-2), one of the catabolic enzymes involved in cartilage degradation, by using semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) technique
Summary
Degenerative joint disease (DJD), often called osteoarthritis, is characterized by the progressive and permanent degradation of articular cartilage [1]. DJD is the most common cause of lameness and poor performance in equines leading to the reason for culling or early retirement from riding in sports horses [3] This disease is the result of the metabolic imbalance of cartilage matrix biomolecules which appears in the degradation exceeding synthesis [4]. A recent study in IL-1β-treated human chondrocytes found that this compound inhibited the expression of MMP-1, MMP-3, and MMP-13 while increasing the mRNA expression of the natural inhibitors of these enzymes is called tissue inhibitors of matrix metalloproteinase (TIMP) via the NF-κB signaling pathway [12] These studies led us to suspect that this effectiveness of andrographolide may implicate protective activity against cytokine-induced cartilage degradation in equine. The results of this study could provide evidence of the chondroprotective potential of A. paniculata for alternative use in horses with DJD
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